姜黄素抑制HeLa细胞诱导的血管内皮细胞增殖作用及其机制研究

Effect and mechanisms of curcumin on proliferation of vascular endothelial cells induced by HeLa cells

目的:研究姜黄素对HeLa细胞诱导的血管内皮细胞增殖的抑制作用及其机制。方法:采用台盼蓝排染法计数人脐静脉内皮细胞系ECV304细胞,MTT法测定不同浓度姜黄素对ECV304细胞和HeLa细胞诱导的ECV304细胞增殖的抑制;采用RT-PCR和Western blot检测40μmol/L姜黄素作用于HeLa细胞诱导的血管内皮细胞4、8、16和24h后,血管内皮生长因子(VEGF)及其受体VEGFR2(KDR)基因及蛋白的表达。结果:姜黄素直接作用于ECV304细胞,其细胞计数较对照组显著减少(P<0.05),并呈浓度依赖性。MTT法测得不同浓度姜黄素作用后,可产生显著的细胞增殖抑制作用,48h体外半数抑制浓度(IC50)值为(38.4±0.031)μmol/L。随着姜黄素浓度的增加,其对ECV304细胞和HeLa细胞诱导的ECV304细胞的增殖有明显的抑制作用,P值分别为0.0000和0.0001。姜黄素作用于HeLa细胞诱导的血管内皮细胞4、8、16和24h后,能明显下调VEGF与KDR的mRNA和蛋白表达。结论:姜黄素能直接抑制血管内皮细胞的增殖,对HeLa细胞诱导的血管内皮细胞的增殖也有抑制作用。此作用可能是通过抑制VEGF及其受体KDR的mRNA和蛋白表达来实现的。

OBJECTIVE： To study the suppressive effect and its mechanisms of curcumin on growth and proliferation of vascular endothelial cells induced by HeLa cells. METHODS： The taipanlan stain was used to count the number of HUVEC cell line ECV304 cells and MTT essay was used to examine the suppressive effect of curcumin on the proliferation of ECV304 cell and HeLa cell induced ECV304 cells. Half-quantitative RTPCR and Western blot assay were used to determine the expressions of gene and protein of VEGF and VEGFR2 （KDR） after 40 μmol/L curcumin interacted with the vascular endothelial cells induced by HeLa cells. RESULTS： Curcumin directly acted on the ECV304 cells. The cell number and proliferation rate were reduced significantly compared with the control group （P〈0.05） in a dose-dependent manner. IC50 of curcumin in ECV304 was （38.4±0.031） μmol/L at 48 h. Curcumin obviously suppressed the proliferation of vascular endothelial and HeLa cell induced vascular endothelial cells in a dose-dependent manner, and P were 0. 000 0 and 0.000 1, respectively. Curcumin obviously down-regulated the expressions of mRNA and protein of VEGF and KDR at different time （4, 8, 16 and 24 h）. CONCLUSIONS： Curcumin can suppress the proliferation of vascular endothelial cells directly, and also has the suppressive effect on proliferation of HeLa cell induced vascular endothelial cells, which are acted by down-regulation of the expressions of mRNA and protein of VEGF and KDR.